text:
EVERY COMMUNITY NEEDS A DAVID CARPENTER
Your humble typist was blessed to have David Carpenter as a partner for several years, primarily in the 1990s in Independent Baseball franchises in Thunder Bay, Ontario and in a greater way in Waterbury, Connecticut. The Waterbury team was known as the Spirit, then of the Northeast League and later the Eastern Division of the Northern League. It might as well have been named the Spirit for David, because he certainly had it.
Mr. Carpenter, who was an all-too-young 69, was laid to rest on a tranquil hillside near his Woodhull, NY farm Monday.
Every community with an Independent Baseball team--or desiring one--should have a David Carpenter. Unfortunately, not nearly enough exist.
He was a man of immense, self-made wealth, which is very important to pay the bills, especially if things do not go well in a baseball startup. The wealth came from his brilliance in the field of zinc oxide applications, where he helped build a Fortune 500 manufacturing company and where he was known as Dr. Carpenter.
But what made David Carpenter special was his nature. He was an uncommon man with a commoner's touch. The glass was always brim full, at worst. It was never half empty. He saw the positives in everything and in every person, as was pointed out so poignantly by those who spoke during the touching funeral service. He treated everyone exactly the same, with an attentive ear to hear his or her story. He did not step forward to tell his story. It was yours that was important.
David Carpenter was so modest he was not a high profile person during his years in Independent Baseball, where he could have offered so much more if only he had been asked. He would have impacted still more lives.
Subscribe now for 2007 Independent Baseball Insider columns...price reduced to $25
Posted by Bob Wirz at Tuesday, June 26, 2007
MEETING THE PIRATES' ED OTT, IN A DIFFERENT ENVIRO...
BOB GIBSON'S SON CHRIS HELPING GATEWAY BUILD A NIC...
THIRD NEW FRANCHISE SET FOR 2008
BUD SMITH AND LONGTIME WOMEN'S STAR TAMARA IVIE WI...


Me and my husband just came back from Chung Hwa, Likas. We just registered Sean in Chung Hwa primary school for 2010. Once i stepped in the school compound, it reminds me of my schooling time….mix feeling.. excited, scared, nervous.. all in one. I am there as an adult.. register for my son.. meaning i'm old lah. I feel relief once i completed the form and given a slip 'Pendaftaran Masuk ke Darjah Satu', the clerk requested me to call the school on November 2009 for confirmation, wah.. i hope i wont forget.
I was reading my favourite magazine 'Women's Weekly' and there was an article about baby and children which feature few style and brands of baby cribs. It suddenly reminds me how grown up my Sean is.. he is no longer baby oh..
I cant believe it he's going to primary school, even now he is in his first year kinder, i just cannot imagine looking at him with the white and blue uniform. Another 2 years.. he will be in Primary School.. wah.. I cant believe it.. and i cant wait.
he he sia pun so excited when Shania started Primary 1! uniform biru bah.. he he i brabis take photo lagi. Our babies grow so fast, don't they?
Have a nice day, Shirley.. take care!
that remind me to register Josh, aduh!
so funny oh.. when you told me that you want to register Amber for primary one..sya trus confuse oh.. hehe. because Sean is older than Amber.


If you have a Pinterest business account then chances are the new Pinterest Showcase feature has shown up on your profile page.
This new feature means when someone visits your profile on Pinterest, you can show them a hand-picked selection of what you have to offer. As you can see it sits right at the top of your profile so it is the first thing people see. It is animated and rotates through up to five of the boards you choose to feature.
This gives you the ideal opportunity to introduce yourself and give a great snapshot of your business and products. If you happen to have 'buyable' pins you can showcase those in the first 'Shop' slot (if you are in the U.S. and have 'buyable' pins these will show up automatically in your shop tab). You are also able to update your showcase as often as you like so you can highlight items that are on special, in season or popular.
If you are a blogger you will want to showcase your blog board so people can see immediately what you are all about. After that, you might want to choose boards that are related to the topics you blog about or even add in 1 or 2 of your favorite personal boards to show off a bit of your personality so you can connect with like-minded followers.
If you sell products, either as a small shop owner or perhaps a direct seller, you are going to want to feature boards that showcase your products! As a direct seller, you can create pins that link back to your replicated site or even to your other social media channels where people can contact you to purchase your products.
Again your other featured boards should somehow link with your main business focus, such as ways to use your products or showcase your own brand personality. You want to entice new visitors to your profile to stick around and follow your boards.
Check out these new showcases from Burberry and mine!
There has been a bit of controversy going around in the Pinterest community with all of the layout changes that have been occurring lately. Pinterest has made these changes so that the 'look' of Pinterest is more consistent across all devices. Everything should now look consistent no matter where people choose to view your profile.
Have you loaded up your showcase yet? What do you think of the latest update – do you think it will help you feature your business better? Let us know in the comments!
Hi! Thanks for this overview. I set up my showcase when it first launched but it hasn't been strategic AT ALL. This gives me some ideas to explore!
That's great I'm so glad it sparked some ideas!


I have seen tons of British, American, Australian kids coming back from CMS to the ferries (mainly Discovery Bay) so guessing there is a lot of English spoken outside of class.
Depending on the age, kids can pick up Chinese BUT...homework, yeah, that can be an issue as you can't really help with that. I see the kids from the Peng Chau school on the ferry going on field trips and the Filipino kids often sit together and speak English with each other.
There is also a school that teaches mostly in English but is designed for learning Chinese- from the names of the kids (they post their work) seems to manly be Nepali kids but some British, American, and Filipino names. It is less well known/popular might be easier to get a spot (because you know about all those issues with primary school spots, I am sure).
By the way, depending on where you work (Central, Sheung Wan, Wan Chai?) Peng Chau is a doable commute and many schools have buses that pick up at the ferry pier. Lots of kids "commute" to school (even kindergarten kids!). It's a nice, healthy lifestyle here, kids riding bikes everywhere, running around and playing, more normal life than other places in HK. The local school teaches English well so even the Chinese kids are pretty good- plus their practice with the Filipino kids. Maybe it wasn't somewhere you were thinking about but check out all the islands area (Mui Wo has a decent-sized Filipino population but haven't seen as many kids at their church as at our church). Basically I am the only non-Filipina at my church but people are very warm and welcoming.
Thanks for sharing. Work will be in Queen's Road Central. I saw that the commute to Peng Chau or Discovery Bay is 1+hour each way. Might be too stressful for a newcomer to HK. May consider in the future as housing in Discovery Bay is way better and more value for money than on island.
DB is 25 minutes by ferry each way, Peng Chau 30 mins or 40 mins depending on which ferry you take. Relaxing boat ride, then just an easy walk. PC, everywhere is 5-10 mins to ferry pier (most people bike there or walk if they live close). DB, well I guess most bus to the pier.
Depending on where you live in HK island, commute time can be about the same. And longer if you live on Kowloon.
For kids, well, might be longer commute depending on bus ride. Not sure if the buses have a lot of stops. But seems a lot of kids do it so there are other kids to hang out with. Depends really on the child- some children want a lot of outdoor space, others don't care (my own daughter had a lot of outdoor space but was a bookworm who couldn't have cared less...).
Good luck in the search, I have a coworker just doing all the kindergarten applications and it's more work than my daughter did applying to university!!
Mine would be an incoming Primary 4 DOB 31 Dec 2010. Hopefully it wont be as tough. Will definitely consider your points, very helpful. Hope to catch up with you soon in HK.


text:
Part of Energy Management Network »
Load Management Group
In partnership with PLMA, this group is for practitioners from energy utilities, solution providers, and trade allies to share load management expertise and explore innovative approaches to program delivery, pricing constructs, and technology adoption.
Emerging Need for Demand Side Flexibility In Our Gas Networks
image credit: istockphotos.com
Donald Mcphail 651
Senior Manager, Product Marketing Uplight
Donald McPhail is a leader and senior specialist with a passion for strategy development and driving transformational change with experience in Australia, US, UK and Netherlands. He has the rare...
15 items added with 5,981 views
Mar 9, 2021Mar 9, 2021 10:32 pm GMT
The concepts of controlling customer smart thermostats or nudging customer behaviors to shift loads outside of peak demand periods are heavily utilized in the management of the electric grid. Yet residential load management - or demand response (DR) - is a tool that historically has not been utilized in the management of supply of natural gas. But that looks to be changing.
The Underexplored Opportunity
Almost as if they spurred on the market, the Brattle Group released a study in 2018 arguing that polar vortex events and the stress they place on natural gas grids - typically resulting in wholesale price spikes - presented an opportunity to leverage DR as part of the solution. In particular, they highlighted gas DR has the potential to:
Help reduce sharp increases in gas and electric price spikes in the short-run and improve electric sector reliability (which relies upon gas for peaking generators)
Drive long-term value by deferring or avoiding costly gas grid investments
Provide environmental benefits by displacing oil in electric generation by freeing up cleaner natural gas, and in the long term by avoiding a natural gas "lock-in"
Rather than relying on adhoc behavioral interventions, the Brattle study highlighted that the industry could expect even better results when coupled with devices such as smart thermostats, which allow for remote control of gas furnaces–– ultimately creating a more dependable residential DR resource. In fact, researchers at the National Renewable Energy Laboratory (NREL) and US Department of Energy found that gas DR with residential smart thermostats could reduce demand by 1-14% during extreme cold-weather conditions.
A Growing Need
By 2023, New York utility ConEd estimates it will only be able to meet 78% of its peak-day demand, based on firm pipeline contracts. Given regulatory challenges around adding pipeline capacity, ConEd has started looking at alternative measures like DR to help ensure supply in the future. As a result, in 2018 the NY Public Service Commission approved ConEd to run a 3 year residential gas DR pilot program with smart thermostats.
The North American Electric Reliability Corporation (NERC) advised that "extreme weather could result in natural gas shortages and challenges to grid reliability in New England and California" due to reliance on natural gas powered peaking generators this winter. The challenge is further exacerbated by the uncertainty of the pandemic on growing demand, and the lack of storage and redundancy in natural gas supply infrastructure.
Learning From Trailblazers
Responding to backbone pipeline constraints and reduced availability of its Aliso Canyon facility, Southern California Gas launched North America's first residential gas DR program in 2017. Growing to more than 44,000 enrolled customers, the program targets short-term 4-hour events by dispatching temperature setbacks on customer smart thermostats. While the program has been credited with proving smart thermostats can be used to reduce gas demand during targeted periods of time, Nexant has highlighted that the "snapback" effect can significantly undermine daily savings. Unlike in the electric system where peak demand relief is typically only required for 2-4 hours, constraints in natural gas infrastructure can typically last for 24 hours. Therefore, while gas consumption was reduced by 15% during the 4-hour event window, the daily savings were only 1-2%. During winter 2019/2020, ConEd also found the snapback effect to be problematic, reducing event savings by 52% on average.
Off the back of its successful summer electric smart thermostat DR program, Consumers Energy launched a gas DR pilot in winter 2020/2021 with 3,000 residential customers. The program will test both short-duration (4-hour) DR events like those run by ConEd and SoCalGas, as well extended (24-hour) DR events. The latter specifically draws on Uplight's experience of running daily "energy efficiency" (EE) optimizations with its Orchestrated Energy platform to save 9 to 16% of heating energy use in a home, on top of the deemed EE savings of smart thermostats. By learning individual customers' comfort preferences and the thermal capacity of a home through normal operation of the smart thermostat, this approach allows for maximizing gas demand reduction for the duration of day-long events, while maintaining customer comfort to avoid overrides and dissatisfaction.
Will it Spark or Fizzle?
While still in its infancy, there is no doubt that residential gas DR shows a lot of potential for the industry - particularly in regions with current or emerging pipeline or storage constraints. Early pilots have also shown that customers are willing and interested in participating. There is also the potential for dual fuel utilities to stack energy efficiency rebates alongside electric and gas DR participation incentives to reward customers for participating, as well as improve the cost effectiveness of each.
As natural gas utilities and regulators look to evaluate the potential for gas DR in their region, they should:
Assess the gas supply curve and contingency scenarios (e.g. one in ten years) to determine the regional need, potential, cost, and timing of gas DR to allow for incorporating infrastructure planning and comparing against alternative solutions
Assess regional policy and regulations that may inadvertently be creating a barrier or disincentive to natural gas utilities running residential DR programs (e.g. disproportionately incentivizing capital investments over demand side management alternatives)
Work with industry - including aggregators, third-party evaluators, smart thermostat manufacturers, and smart water heater controller manufacturers - to inform program design and implementation strategies based on best practices and insights from other programs
Continue working with experienced residential DR solution providers to pilot, evaluate and scale programs
While the use of residential DR for gas networks is new, there is clearly a growing need to manage limited gas resources. While it might seem overwhelming for asset managers on where to start, looking to existing DR gas programs can help a utility start small, test and learn, and then build up their capability. By doing so the natural gas industry can follow the path of their electric cousins, and create a true win-win for utilities and customers alike.
customer-experience-&-management
demand-management
demand-response
energy-efficiency
natural-gas-generation
Donald Mcphail
Matt Chester on Mar 9, 2021
Do you think part of the reason it's underexplored is because it might be perceived as 'getting in the way' of electrification efforts that would render gas flexibility a moot point? Might it also risk painting gas in a 'greener' light in a way that will keep the fuel around longer than it may have otherwise?
Donald Mcphail on Mar 9, 2021
I think the reason for being underexplored historically is that the challenges of resource adequacy is different in gas networks compared to electric, and typically gas utilities haven't been short on capacity. The challenges in SoCal on reduced supply or NY with restrictions on distribution infrastructure expansion are more recent and local challenges, that aren't a problem in say the midwest. Where it does get really interesting is the intersection of gas as a fuel for electric supply vs for end user heating. That is an area where I can see gas DR playing a valuable nearer term role.
I wouldn't say that full electrication of heating in the very near real term is a given. There are certainly regions where cannibalization of gas customers into all electric isn't allowed, and all gas utilities don't have a value proposition to convert the customers to electric. But regardless demand side flexibility of gas can certainly help with transitions and resiliency, whether it be to avoid new supply or pipeline additions, or to compliment winter supply peaks in electric system. It's even more attractive when the same resources are being used to provide electric DR in the summer and gas DR in the winter.
Matt Chester on Mar 10, 2021
But regardless demand side flexibility of gas can certainly help with transitions and resiliency, whether it be to avoid new supply or pipeline additions, or to compliment winter supply peaks in electric system. It's even more attractive when the same resources are being used to provide electric DR in the summer and gas DR in the winter.
Great point, Donald-- thanks for your detailed reply! I'll be eager to see if the idea of gas demand flexibility takes hold in the near term like it's started to do for electricity
Thank Donald for the Post!
Maximizing Value from DERs Through Value Stacking
Managing the Impacts of COVID-19 on Power and Energy with Demand Response
Set it and Save: Using Technology to Ensure a Smooth TOU Rate Transition

redpajama_set_name: RedPajamaCommonCrawl


text:
Brooklyn's A.I.R Gallery to Launch Its First Feminist and Queer Art Book Fair
The veteran feminist, artist-run nonprofit will offer a selection of books by publishers that prioritize feminist and queer histories.
by Hakim Bishara January 6, 2020 January 3, 2020
A vintage issue of M/E/A/N/I/N/G on display at the Pratt Institute Libraries (courtesy of Mira Schor and Susan Bee)
A.I.R. Gallery, a nonprofit organization in Brooklyn, will soon launch its first Feminist & Queer Art Book Fair. Participating publishers and book artists will include Belladonna, Dancing Foxes Press, GenderFail, Litmus Press, MOMMY, Pinsapo Press, Precog Magazine, Siglio Press, Visual AIDS, and A.I.R. artists among others.
"The fair aims to offer space for artistic experimentation, prioritize feminist and queer histories, and facilitate intergenerational dialogue," A.I.R. explains. "The gathering of publishers across media and subject prompts community-building and knowledge sharing, while celebrating the long history of A.I.R. artists who have been bookmakers working in language and form."
Established in 1972, A.I.R. Gallery was one of the first women-led, artist-run, nonprofit organizations dedicated to women artists in the United States. The gallery operated at 97 Wooster Street in Soho before moving to 111 Front Street in Brooklyn's Dumbo neighborhood in 2008. In 2015, A.I.R. moved to its current location at 155 Plymouth Street. One of the gallery's most celebrated initiatives is the Fellowship Program For Emerging and Underrepresented Women Artists, which provides a year-long career development intensive for six artists annually, as well as life-long support and collaboration.
Highlights at the fair will include books and cards from the 1980 exhibition "Speaking Volumes" curated by Lucy Lippard for A.I.R. — featuring more than 100 books by women-identified artists — as well as Susan Bee and Mira Schor's collaborative project, M/E/A/N/I/N/G magazine.
The fair is free to participants and the public alike. A.I.R. Gallery is wheelchair accessible via a ramp. There are accessible toilets in the venue.
When: January 18, 2020, 12-7 pm
Where: A.I.R. Gallery, 155 Plymouth Street, Brooklyn NY
More info at A.I.R. Gallery
Join Residency Unlimited for Panels on Feminist Activism and Leadership in Visual Arts
Art Books for Days: What Not to Miss at Printed Matter's 2021 Virtual Art Book Fair
Tagged: A.I.R. Gallery, Events, New York
Hakim Bishara
Hakim Bishara is a staff writer for Hyperallergic. He is also a co-director at Soloway Gallery, an artist-run space in Brooklyn. Bishara is a recipient of the 2019 Andy Warhol Foundation and Creative Capital...	More by Hakim Bishara
One reply on "Brooklyn's A.I.R Gallery to Launch Its First Feminist and Queer Art Book Fair"
sabjonze says:
The editors of "Shameless Feminists" from World War 3 Illustrated are looking forward to being a part of this event!


Well my name is Johnny, I hope to get to know the people on this site well. I haven't Rped since I graduated Highschool so I may be less then stellar, but i'm willing to give threads a go if you all are willing to deal with the rusty posts.
P.S. - Your register button on the home screen is broken I believe, it kept redirecting me to the home page even on multiple computers and an iphone.
Hey, Johnny! Welcome to HBH! I'm sure you'll do great. Looking forward to RPing with you!
Hey Johnny! Hope you like it here!
Hope you enjoy RPing here Johnny!
Glad you're excited Joseph, I hope you enjoy your time with us!!!
thanks. please feel free to call me Joe.
Don't hesitate to hit any of us up if you wanna rp.
A little late to jump on the train, but welcome, Joe!
Hi, I'm Alexandria. I think it's pretty obvious that I'm new. My character, Ada Eloway, is a 14-year-old Roman demigod. I don't know how to sum her up right now, so I suggest reading her biography. It's short compared to a lot of the ones I've read around here. I haven't role-played in a few years, so I'm out of practice. Hopefully I can relearn quick enough. I'll probably be slow in getting started. My job is moving me across the country this week. The process is weird for my situation, so I do not know how much I will be here. I hope to interact with you all and find my niche soon.
Welcome to HBH, Alexandria! Glad to have you here, looking forward to RPing with you!
Hey there, I'm Raven(or my online alias is). I'm new here and really excited to RP with you guys.
Welcome onto Half Blood Hill RPG Raven, I hope you have a nice stay!
Hey Raven, welcome to HBH!
Feel free to PM anyone here if u wanna plot.
Welcome to HBH, Alex! Glad to have you here, looking forward to RPing with you.
Heya, im River and im really weird!!


The possibility that autologous NK cells could serve as an effective treatment modality for solid tumors has long been considered. However, implementation is hampered by (i) the small number of NK cells in peripheral blood, (ii) the difficulties associated with large-scale production of GMP compliant cytolytic NK cells, (iii) the need to activate the NK cells in order to induce NK cell mediated killing and (iv) the constraints imposed by autologous inhibitory receptor-ligand interactions. To address these issues, we determined (i) if large numbers of NK cells could be expanded from PBMC and GMP compliant cell fractions derived by elutriation, (ii) their ability to kill allogeneic and autologous tumor targets by direct cytotoxitiy and by antibody-mediated cellular cytotoxicity and (iii) defined NK cell specific receptor-ligand interactions that mediate tumor target cell killing.
Human NK cells were expanded during 14 days. Expansion efficiency, NK receptor repertoire before and after expansion, expression of NK specific ligands, cytolytic activity against allogeneic and autologous tumor targets, with and without the addition of chimeric EGFR monoclonal antibody, were investigated.
Cell expansion shifted the NK cell receptor repertoire towards activation and resulted in cytotoxicity against various allogeneic tumor cell lines and autologous gastric cancer cells, while sparing normal PBMC. Blocking studies confirmed that autologous cytotoxicity is established through multiple activating receptor-ligand interactions. Importantly, expanded NK cells also mediated ADCC in an autologous and allogeneic setting by antibodies that are currently being used to treat patients with select solid tumors.
These data demonstrate that large numbers of cytolytic NK cells can be generated from PBMC and lymphocyte-enriched fractions obtained by GMP compliant counter current elutriation from PBMC, establishing the preclinical evidence necessary to support clinical trials utilizing autologous expanded NK cells, both directly and in combination with monoclonal antibodies in future cell-based immunotherapy in select solid tumors.
Natural killer cells (NK) were identified more than 30 years ago as a population of lymphokine activated killer cells that showed the ability to kill tumor cells in vitro in the absence of prior immune sensitization of the host [1–4]. Over the ensuing years, much has been learned about regulation of their biologic activity and, in particular, their potential use as an immunotherapeutic modality in cancer .
It has become clear that the biologic activity of NK cells is controlled by a complex repertoire of surface receptors which, upon engagement by ligands on a target cell, signal either an inhibitory or activating response . The major inhibitory and activating receptors are products of germ line genes encoding killer cell immunoglobulin-like receptors (KIRs) and in an autologous environment, inhibition of NK cell cytotoxic activity is dominant and governed by epitopes on self HLA class I alleles. In general, cytotoxic activity of NK cells is triggered when the target cell lacks expression of some or all HLA class I molecules; the basis for the "missing self" hypothesis .
Recognizing the possibility that NK cells have the ability to kill tumors that lack expression of the inhibitory HLA class I alleles, investigators have reported significant antitumor responses in clinical settings of allogeneic stem cell transplantation. Importantly, clinical effects are demonstrated when inhibitory effects are bypassed by utilizing haplo-identical NK cells and best results are achieved when, in addition, KIR-ligand mismatched NK cells are selected [8, 9]. In turn, this approach requires extensive donor screening and careful depletion of allogeneic T cells from the NK cell product before administration to the host in order to avoid the risk of graft-versus-host disease (GvHD) .
The possibility that infusion of autologous NK cells could serve as an effective treatment modality for solid tumors has long been considered . However, implementation is hampered by (i) the small number of NK cells in peripheral blood that could be isolated relative to the number of cells that would be required to be effective and the difficulties associated with large-scale production of cytolytic NK cells in compliance with Good Manufacturing Practices (GMP), (ii) the need to activate the NK cells in order to induce NK cell mediated killing of a resident tumor and (iii) the constraints imposed by autologous inhibitory receptor-ligand interactions.
The first issue has been addressed in a number of reports that demonstrate that large numbers of NK cells could be expanded from CD56+ cells isolated from peripheral blood mononuclear cells (PBMC) obtained from healthy individuals and patients with hematological malignancies and solid tumors. Expansion was achieved by short term culture with cytokines alone, by cytokines and co-culture with irradiated feeder cells consisting of EBV transformed lymphoblastoid cell lines or cytokines and co-culture with K562 cells that had been transfected with and expresses cell membrane-bound IL-15 and 4-1BBL [12–16]. In most instances, these expanded cells were generated from NK cells (CD56+CD3-) isolated from peripheral blood using magnetic beads. The expanded NK cells were highly cytotoxic when tested against variety of target cells that consisted primarily of allogeneic cancer cell lines established from hematologic malignancies [12, 17].
In addition, a GMP compliant and closed system has successfully been established for the enrichment of monocytes from PBMC using counter current elutriation . Besides a highly enriched population of monocytes, lymphocyte-enriched fractions are also obtained. Currently, clinical studies are ongoing utilizing elutriation derived monocytes for large-scale generation of dendritic cells in order to treat a variety of metastatic cancers.
The objectives of this study were to evaluate if the aforementioned strategies could be combined in order to expand large numbers of NK cells from PBMC from normal individuals and patients with various solid tumors. Furthermore, the possibility to expand NK cells from lymphocyte-enriched cell fractions derived from PBMC by elutriation rather than utilizing isolated CD56+ cells as the starting cell population was determined. In addition, cytolytic allogeneic and autologous activity by direct cytotoxicity as well as antibody-mediated cellular cytotoxicity and NK specific receptor-ligand interactions that mediate target cell killing were defined.
We confirmed that large quantities of cytotoxic NK cells can be expanded from PBMC in the presence of K562 cells expressing membrane-bound IL-15 and 4-1BBLigand from normal individuals and patients with various solid tumors. Ex-vivo expansion tended to alter the balance of NK cell receptor expression towards those that activate and mediate cytotoxicity. This activity resulted in cytotoxicity against various allogeneic tumor targets and more importantly, against autologous-derived gastric tumor targets. Blocking studies identified multiple activating receptor-ligand interactions that would be predicted to mediate NK cell cytotoxicity. Moreover, these activating receptor-ligand interactions were operative in antibody-dependent cellular cytotoxicity (ADCC) in an allogeneic and autologous setting. Importantly, as a mean for future clinical translation, GMP compliant cytolytic NK cells could efficiently be expanded from lymphocyte-enriched cell fractions obtained from PBMC by counter current elutriation.
Our studies demonstrate that human NK cells acquire cytolytic activity against autologous gastric tumor cells after ex-vivo expansion and suggest a therapeutic potential for autologous expanded NK cells, both directly and in combination with monoclonal antibodies in future cell-based immunotherapy.
Human blood samples were purchased (BRT Laboratories, Baltimore, MD) and whole peripheral blood mononuclear cells (PBMC) were isolated using density-gradient centrifugation. Using leukapheresis products purchased from the same source, the constitutive cell populations were fractionated by continuous-counterflow elutriation following protocols established by the manufacturer of cell separator (Elutra, Gambro BCT). This instrument uses continuous counter-flow elutriation technology to separate cells fractions based primarily by size and secondarily by specific gravity. In brief, the leukapheresis product was loaded via an inlet pump into a constantly rotating (2400 rpm) elutriation chamber. Based on centrifuge speed and cell density, five elutriated cell fractions were collected. PBMC and various elutriated cell fractions were viably frozen in RPMI-1640 (Invitrogen Corp., Grand Island, NY) supplemented with 20% human AB serum (Gemini Bio-Products, Woodland, CA) and 10% Dimethylsulfoxide (Sigma, St. Louis, MO) using an automated cell freezer (Gordinier Electronics, Roseville, MI) and stored in the vapor phase of liquid nitrogen until used.
The myeloid cell line K562, prostate cancer cell lines LNCaP, PC-3 and DU-145 and breast cancer cell line MCF-7 were available in our laboratory. The lung cancer cell line H358 was kindly provided by Dr. S. Ostrand-Rosenberg (Department of Biological Sciences, University of Maryland Baltimore County, Catonsville, MD) and the Head and Neck cancer cell line TU-167 was kindly provided by Dr. S. Strome (Department of Otorhinolaryngology-Head and Neck Surgery, University of Maryland, Baltimore, MD).
With patient consent and under approval of the Institutional Review Board, peripheral blood mononuclear cells were obtained from 2 patients with gastric cancer undergoing treatment at the Tokyo Clinic and Research Institute. Cell lines (tumor 1 and tumor 2) were established from biopsies of metastatic gastric tumor lesions from the respective patients. All tumor cell lines were cultured in RPMI 1640 supplemented with 10% Fetal Bovine Serum, 1% P/S and 1% Glutamax-1 (cRPMI).
NK cells were expanded from PBMC as previously described with some minor modifications . In brief, PBMC (1.5 × 106) were incubated with irradiated (14,000 rad) K562-mbIL15-41BBL cells (106) in a 24-well tissue culture plate in the presence of 200 IU/ml human IL-2 (R&D Systems Inc) in cRPMI. Half of the culture medium was replaced every 2-3 days with fresh culture medium for the first 6 days. After 6 days of expansion, cells were harvested, washed, counted and re-cultured at a starting cell density of 1 × 105-3 × 105/ml in T-25 or T-75 culture flasks in cRPMI supplemented with IL-2. Cells were expanded for and additional 8 days. Additional cRPMI was added to the flasks if necessary based on cell density.
Cell surface expression was determined before and after 14 days of cell expansion by staining with directly conjugated mouse anti-human mAb's against CD3, CD56, αβTCR, γδTCR, HLA class I, HLA-DR, Fas, Fas-ligand, KLRD1, NKG2a, KIR3DL1, ILT2, CD62L, KIR3DL2/3, NKG2d, DNAM-1, NKp46, NKp44 and NKp30 (BD Biosciences). Gates were set around NK cells which were defined as CD3-CD56+ cells. Surface expression of NK cell ligands was determined on both autologous gastric tumor cell lines and included directly conjugated mouse anti-human nectin-2, PVR, MIC A/B, Fas, HLA class I, HLA class II, HLA-G and purified mouse anti-human HLA-E, ULPB-1, ULBP-2 and ULBP-3. For EGFR-mediated ADCC, gastric tumors were stained with mouse anti-human EGFR mAb. Mouse IgGs were used as isotype controls and purified mAbs were secondarily stained with FITC labelled goat anti-mouse mAb. A minimum of 10000 events were acquired using a BD™ LSR II flow cytometer. Data was analyzed with BD™ FACS DIVA Software.
Cytolytic NK cell activity was measured by 4 hour chromium 51 (51Cr)-release assays as previously described . K562 cells were included as target cells in all cytotoxicity assays to assess overall cytotoxicity performance (data not shown). Expanded day 14 cells were purified into separate populations of NK cells (CD3-CD56+) and NKT/T (CD3+CD56+/CD3+CD56-) cells using MACS human CD3 microbeads and non-expanded NK cells were purified from PBMC using a MACS human NK cell isolation kit. (Miltenyi Biotec Inc). Cell purity was determined to be >92% and 95% respectively. To determine ADCC, 10 μg/ml human IgG1 (huIgG1, Sigma-Aldrich Corp, St. Louis, MO, USA) or Cetuximab (University of Maryland Marlene and Stewart Greenebaum Cancer Center Pharmacy) was added to defined wells. Purified mouse IgG1, mouse anti-DNAM-1, NKp46, NKp44, NKp30 or all four together (all at 10 μg/ml) were added to defined wells during 4 hours of cytotoxicity in order to assess specific activating NK cell receptor-tumor ligand interactions. Reduction in cytotoxicity was calculated based on percentage cytotoxicity in the presence of indicate blocking mAb(s) versus percentage cytotoxicity in the presence of mouse control mAb. The % reduction in ADCC was calculated with percentage cytotoxicity in the presence of human IgG1 set at 100%. To minimize changes that may occur when cell lines are established from primary tumors, the gastric cell lines used in these studies were cultured for less than 10 passages after isolation from the primary tumor tissue.
Paired two-tailed Student's t tests were used to calculate p values. P < 0.05 was considered to be significant.
To achieve large-scale expansion of human NK cells, PBMC were co-cultured in a 1 to 1.5 ratio with lethally irradiated K562 cells expressing membrane-bound IL-15 and 4-1BBLigand (K562-mbIL15-4-1BBL) in culture media containing 200 units IL2/ml. After 14 days of culture, NK cells (CD56+CD3- as defined by flow cytometry) expanded greater than 2 orders of magnitude from PBMC (mean 165 fold; range 4-567 fold, n = 6) and cell products became significantly enriched in NK cells (day 0 with mean 7%, range 3.2%-12.6% versus day 14 with mean 45.6%, range 7.4%-76.4%; P = 0.0140). At the same time, NKT cells (CD56+CD3+ as defined by flow cytometry) expanded at an average of 57 fold (range 7-234), although no significant enrichment (day 0 with mean 3.8%, range 0.8%-8.1% versus day 14 with mean 11.4%, range 2.3%-17.9%; P = 0.1907) was observed. In contrast, a significant decrease in T cells (CD3+ as defined by flow cytometry) was noted after 14 days of expansion (day 0 with mean 54.5%, range 39.9%-71.2% versus day 14 with mean 30.0%, range 4.2%-58.4%; P = 0.0436) with an absolute expansion of 7 fold (range 2-19). The distribution of NK cells and NKT cells in PBMC after expansion is shown in Figure 1A.
Cytolytic NK cells are efficiently expanded from PBMC. In the presence of K562-IL15-41BBL (A) expanded cells become significantly enriched (P = 0.0307) in NK cells (defined by CD56+CD3- cells) after 14 days of culture. Expanded cells were evaluated for cytolytic activity using 4 hour 51Cr release assays. Ex-vivo expanded cells from PBMC (■ donor 1 and △ donor 2), but not freshly purified non-expanded NK cells (◇), efficiently lysed allogeneic tumor cell lines derived from breast (MCF-7) and prostate (LNCaP) cancers but not allogeneic or autologous PBMC derived from donor 1 (B). The mean percentage cytotoxicity is shown from triplicate wells. Error bars represent the SD. Lytic activity is likely mediated by NK cells in the expanded cell population (○) since separation in individual populations of NK cells (◇) and NKT/T cells (△) resulted in allogeneic cytolytic activity of the expanded cell population and the purified NK cell population. Little lytic activity was observed in the presence of NKT/T cells alone (C). The mean percentage cytotoxicity is shown from triplicate wells from one representative experiment. Error bars represent the SD. Experiment shown represents one of three individual experiments with three different donors.
Importantly, ex-vivo expanded NK cells from healthy donor PBMC efficiently lysed allogeneic breast-and prostate-derived tumor targets but not allogeneic or autologous PBMC (Figure 1B). We did observe that cytotoxicity was associated with overall expansion efficiency. Specifically, the one donor whose cells expanded 4 fold after 14 days of culture demonstrated an average of 11.7% cytotoxicity (effector to target ratio 1:10) against K562 cells whereas donors who expanded an average of 202 fold (range 34-576; n = 4) possessed an average of 59.8% cytotoxicity (range 56.0%-65.9%; n = 4) against K562 cells (data not shown).
Based on CD3 and/or CD56 phenotype, the majority of cells in the expanded cell products represented NK cells while a much smaller proportion represented NKT and T cells (Table 1). To determine if both the NK cells and NKT/T cells mediated cytolytic activity, the two populations were isolated by immunomagnetic bead selection and killing assays against prostate-derived tumor cell targets were performed. Cytolytic activity was mediated by NK cells and not NKT cells (Figure 1C). Interestingly, little to no killing was observed with the NKT/T cell population even though a subpopulation of the T cells was confirmed to be γδ-TCR+ by flow cytometry (data not shown). Although γδ-TCR+ T cells are reported to have lytic activity against allogeneic tumor cells, they first require in vitro activation with isopentenyl pyrophosphate (IPP) and IL-2 . Studies are underway to determine if addition of IPP will expand a cytolytic γδ-TCR+ population.
The capacity of K562-mb15-41BBL to stimulate expansion of NK cells from peripheral blood of healthy individuals and children with leukemia in remission was previously demonstrated [12, 17]. However, there is little information in reference to expand NK cells from PBMC derived from patients with solid tumors. With informed consent, NK cells were expanded from PBMC derived from patients with various solid tumors, including gastric cancer, lung cancer, colon cancer and hepatocellular cancer. After 14 days of culture, cell products became significantly enriched in NK cells (day 0 with mean 23.5%; range 5%-46% versus day 14 with mean 80%; range 60%-95%, n = 6, P = 0.0001 data not shown). Expansion efficiency was comparable between PBMC derived from solid tumor patients versus healthy donor PBMC (mean 316 fold; range 1-1795 with n = 6 versus mean 165 fold; range 4-567 with n = 6, P = 0.6685).
These data suggest that NK cells are efficiently expanded from PBMC from normal individuals and more importantly, from patients with various solid tumors without the need of primary enrichment protocols.
Human NK cells maintain self tolerance by the expression of at least one inhibitory receptor specific for autologous HLA class I which prevents cytotoxicity against autologous cells . To establish cytotoxicity against autologous target cells, inhibitory signals must be overcome, either by (i) down-regulation of inhibitory ligands on the tumor cell, (ii) enhanced expression of activating receptors on NK cells, (iii) expression of ligands on the tumor target that activate the NK cell or (iv) a combination of thereof. Since NK cell activation is affected by cytokines such as IL-2 and IL-15 , we sought to determine if NK cells expanded from PBMC were phenotypically different from non-expanded NK cells (Table 2).
In expanded NK cells from normal individuals, no significant change was observed in inhibitory receptors KIR3DL1 (P = 0.1526), KIR3DL2/3 (P = 0.7858) and the activating receptor NKG2D (P = 0.1074). In contrast, activating receptors DNAM-1 (P = 0.0061), NKp46 (P = 0.0161), NKp44 (P = 0.0039) and NKp30 (P = 0.0131) were significantly increased in expression after 14 days of expansion. Interestingly, KLRD1 (P = 0.0012) and NKG2A (P = 0.0118), which both form a complex with the inhibitory non-classical HLA class I ligand HLA-E [23, 24] were also significantly increased after expansion. Importantly, expression of the inhibitory receptor ILT2 (P = 0.0142) which recognizes multiple HLA class I alleles, including non-classical HLA class I, HLA-G , was significantly decreased after expansion.
In order to define the ability of expanded NK cells derived from patients with solid tumors to kill their autologous tumors, tumor cell lines were established from tumor biopsies from two metastatic gastric cancer patients undergoing immunotherapy at the Tokyo Clinic and Research Institute. Of note, the expression of inhibitory and activating receptors on expanded NK cells from the gastric cancer donors were generally not different from expression on expanded NK cells from normal donors (Table 2).
Since autologous NK cell cytotoxicity is the net result of engagement of activating NK cell receptors with activating target cell ligands, the two gastric tumor cell lines were first phenotypically characterized for expression of ligands (Table 3) that are known to engage the NK cell receptors identified in table 2. While the ligands for human NKp46, NKp44 and NKp30 are to be defined, both patient cell lines expressed high levels of the inhibitory ligands HLA class I (75% and 67%, respectively) and HLA-G (42% and 57%, respectively) and relatively small amounts of the activation ligands MHC class I chain-related (MIC) A/B (2% and 1%, respectively), UL16 binding protein (ULBP)-1 (both 3%), ULBP-3 (both 3%) and polio-virus receptor (PVR; 8% and 9%, respectively). Importantly, both cell lines expressed the activating ligand nectin-2 (both 92%; specific for DNAM-1) which prompted us to evaluate both cell lines for their sensitivity against autologous NK cells.
Subsequent 4 hour chromium-release (51Cr-release) assays confirmed that gastric tumor cells derived from both patients were killed by autologous expanded NK cells (Figure 2A) and not by resting (non-expanded) NK cells from patient 2. Unfortunately, insufficient numbers of PBMC from patient 1 were available to isolate and test resting NK cells.
Ex-vivo expanded NK cells recognize autologous gastric tumor cells through different activating receptor-ligand interactions. PBMC from two gastric cancer patients were ex-vivo expanded for 14 days and then tested for cytolytic activity against autologous gastric tumor cells in 4 hour 51Cr release assays. (A) Ex-vivo expanded cells from both patients (patient 1 and patient 2), but not freshly purified non-expanded NK cells from patient 2, efficiently lysed autologous gastric tumor cells (Effector to Target Ratio 20:1). To evaluate the impact of activating receptor-ligand interactions on autologous tumor cell lysis indicated blocking antibodies (10 μg/ml) were added during 4 hours of incubation. (B) Cytotoxicity was reduced in the presence of DNAM-1 (P = 0.0309) and NKp30 (P = 0.0056) for patient 1 and in the presence of NKp46 (P = 0.0003) for patient 2. In both patients autologous cytolytic activity was abrogated in the presence of all four blocking antibodies with P = 0.0111 and P = 0.0001, respectively. Statistical analysis is based on triplicate wells of four (patient 1) and two (patient 2) experiments performed, respectively. Error bars represent the SD. * P < 0.05. MoIgG1 indicates mouse IgG1.
Since expanded NK cells significantly up-regulated DNAM-1, NKp46, NKp44 and NKp30, we performed blocking studies in order to evaluate the importance of these activating receptor-ligand interactions in autologous tumor cell recognition (Figure 2B). As expected, autologous lytic activity was significantly reduced (P = 0.0111 for patient 1 and P = 0.0001 for patient 2) when activating receptor-ligand interactions were interrupted by all four blocking antibodies (mAbs). Specifically, lytic activity of autologous NK cells from patient 1 was significantly reduced in the presence of mAb against DNAM-1 (P = 0.0309) or NKp30 (P = 0.0056) while lytic activity of autologous NK cells from patient 2 was only affected in the presence of mAb against NKp46 (P = 0.003).
Over many years, it has been postulated that eradication of human tumors may best be accomplished by combining cancer treatments modalities [26, 27]. Monoclonal antibodies that react with cell surface structures expressed on cancer cells represent the most successful cancer immunotherapy to date. It is quite clear that their mechanism of action is, at least partially, due to NK cell-mediated ADCC . Since expanded NK cells expressed high levels of CD16 (data not shown), an Fc receptor that mediates ADCC, we sought to determine if lytic activity against the gastric tumor cells could be enhanced in the presence of Cetuximab (Erbitux®), a chimeric monoclonal antibody that reacts with the EGFR receptor and is used to treat patients with a variety of solid tumors .
Both gastric tumor cell lines were screened for EGFR and only one of the two patient tumor cell lines (patient 1) expressed EGFR (Table 2). Subsequent 51Cr-release assays confirmed that allogeneic and autologous cytolytic activity is greatly enhanced in the presence of chimeric anti-EGFR mAb but not in the presence of human IgG1 control antibody (Figure 3A). As expected, the enhancement in cytotoxicity was far more dramatic in the autologous setting if compared to the allogeneic setting since allogeneic tumor cells do not maintain self tolerance through specific inhibitory receptor-ligand interactions.
Cetuximab significantly enhances cytolytic activity and ADCC is negatively affected by inhibition of activating receptor-ligand interactions. Ex-vivo expanded cells from cancer patient 1 were evaluated for their ability to mediate ADCC against autologous (patient 1) and allogeneic (TU-167 and H-358) EGFR expressing lung cancer cells. (A) Cytolytic activity of ex-vivo expanded cells was enhanced in the presence of Cetuximab (10 μg/ml, black bar) but not in the presence of control human IgG1 (10 μg/ml; dotted bar) or media alone (white bar). The mean percentage cytotoxicity is shown from triplicate wells from one representative experiment. Error bars represent the SD. Experiment shown represents one of two individual experiments. (B) The addition of blocking antibodies (10 μg/ml) against DNAM-1, NKp46, NKp44 and NKp30 (= all) significantly reduced (P = 0.0176) Cetuximab-mediated ADCC. Statistical analysis is based on three experiments performed. Error bars represent the SD. * P < 0.05. HuIgG1 indicates human IgG1, Ctx; Cetuximab and moIgG1; mouse IgG1.
Importantly, the expression of activating receptors on the ex-vivo expanded NK cells positively affected overall cytotoxic activity (Figure 3B) since blocking all four activating receptors on the NK cell surface decreased autologous cytotoxicity if compared with control mAb (P = 0.0176 and P = 0.1019, respectively). These data suggest that the combined strategy of adoptively transferred ex-vivo expanded autologous NK cells with infusion of an mAb that is used for cancer immunotherapy may provide clinical benefit for the treatment of select human solid tumors. To extend these observations, we are attempting to establish cell lines from other solid tumors where PBMC would be available to test NK expansion and direct cytotoxicity and ADCC capability.
A GMP compliant system has successfully been established for the enrichment of monocytes from PBMC using an Elutra cell separator. In this closed system, PBMC are fractionated by centrifugal elutriation and five cell fractions are obtained. In general, these fractions consist of platelets (fraction 1), erythrocytes mixed with lymphocytes (fraction 2), lymphocytes (fraction 3), lymphocytes mixed with monocytes (fraction 4) and mainly monocytes (fraction 5) as demonstrated in Figure 4 (n = 11). Current clinical cellular therapy protocols use monocytes obtained from elutriated fraction 5 to generate dendritic cells for cancer immunotherapy while the cells from fractions 2, 3 and 4 are usually "archived" in liquid nitrogen. As a means to facilitate clinical translation, we explored the possibility of these GMP compliant cell fractions to serve in future NK cell-based immunotherapy studies. PBMC and separate elutriated cell fractions were expanded with the aforementioned expansion strategy. After 14 days of culture in the presence of K562-mbIL15-41BBL cells and exogenous IL-2, NK cells expanded greater than two orders of magnitude from PBMC (mean 165 fold; range 4-567 fold with n = 6, data not shown), elutriated cell fraction 2 (mean 209 fold; range 3-615 fold with n = 3, data not shown), elutriated cell fraction 3 (mean 131 fold; range 4-339 fold with n = 3, data not shown) and elutriated cell fraction 4 (mean 91 fold; range no expansion-358 fold with n = 4, data not shown). Importantly, expanded cells from PBMC and separate elutriated cell fractions became significantly enriched in NK cells and lysed allogeneic prostate-derived tumor cell lines in a similar fashion (Figure 5A-B). Thus, these data show that large quantities of cytolytic NK cells can be expanded from various elutriated cell fractions collected with the GMP compliant Elutra system.
Distribution of lineage-specific phenotypic markers on PBMC and separate cell fractions obtained after counter current elutriation. PBMC and elutriated cell fractions were stained with various lineage-specific directly-conjugated antibodies and analyzed by flow cytometry (A). Average number of cells and phenotypic distribution (%) expressing lineage-markers in elutriated cell fractions (n = 11) (B).
Ex-vivo expanded cells from elutriated cell fractions efficiently lyse allogeneic prostate cancer cells. PBMC and elutriated fractions 2, 3 and 4 from the same healthy individual were expanded ex-vivo in the presence of K562-mbIL15-41BBL and IL-2 for 14 days and then tested for in vitro cytolytic activity. Cytolytic activity was evaluated in 4 hour 51Cr release assays against (A) prostate cancer (DU-145, PC-3 and LNCaP) cells. Ex-vivo expanded cells from elutriated cell fractions 2 (◇), 3 (△) and 4 (□) lysed prostate cancer cells in a similar fashion as ex-vivo expanded cells from PBMC (○). (B) Elutriated cell fractions become enriched in NK cells (defined by CD56+CD3- cells) after 14 days of culture regardless the cellular content of these fractions. The mean percentage cytotoxicity is shown from triplicate wells from one representative experiment. Bars represent the SD. Experiment shown represents one of four individual experiments.
The use of NK cells as a cancer treatment modality in the absence of allogeneic stem cell transplant requires that large quantities of NK cells are generated that kill the tumor cells directly or augment the cytotoxic effect of tumor directed monoclonal antibodies. The critical findings of the studies reported herein are that (i) the technology that was reported to successfully expand NK cells isolated from PBMC [12, 17] was also effective in generating large numbers of NK cells from PBMC and various elutriated cell fractions without previous isolation of the CD56+ cells, (ii) activation receptors and other cell surface structures that mediate target cell killing are increasingly expressed on the expanded NK cells, (iii) NK cells expanded from PBMC from patients with solid tumors kill allogeneic and autologous cancer cell lines by direct cytotoxicity and, importantly, mediate ADCC to autologous tumor cell targets in the presence of chimeric mAb and (iv) the interactions of activating receptors DNAM-1, NKp30 and NK-p46 with cognate ligands on the tumor target appear to mediate direct and ADCC mediated autologous cytotoxicity in gastric carcinoma.
For translation into the clinic it is important to observe that besides NK cells, relatively small numbers of NKT and T cells are expanded in this system. These cell populations may mediate GvHD when infused together with NK cells in adoptive allogeneic immunotherapy protocols. GvHD is a serious, potentially life-threatening, condition resulting from transplanted or infused allogeneic donor cell recognition of the recipients' tissues as non-self, and is predominantly mediated by CD3+ T cells . These cells are often depleted to prevent GvHD, as could be accomplished with the cells expanded by the protocol presented here. Depletion of T cells from the NK cell product before administration to the host is likely to be less critical in the autologous setting.
An important observation in our studies was that the expanded NK cells did not kill autologous and allogeneic PBMC, an indication that despite the increase in surface expression of activating receptors on the NK cells, the inhibitory ligands expressed on normal PBMC were dominant and able to control cytolytic activity against non-malignant cells. This is further illustrated in that both gastric tumor cell lines were susceptible to autologous cytotoxicity despite the expression of high levels of inhibitory classical and non-classical HLA class I molecules. These data suggest that, under certain conditions, activating receptor-ligand recognition may override receptor-ligand interactions that inhibit NK activity. Emerging data indicates that important triggers in this interaction are surface structures (ligand) that are expressed on cells that have undergone malignant transformation. In addition, it is well recognized that HLA class I expression the major NK cell inhibitory structure, is often down regulated in many solid tumors. In the case of autologous NK cell cytotoxicity against PBMC, inhibitory signals still predominated over activating signals, since no cytotoxicity of NK cells against autologous or allogeneic PBMC was observed. Our results indicate that the NK cells expanded and activated by the methods described do not recognize and kill non-transformed cells.
In addition, while significantly higher levels of the inhibitory CD94/NKG2A complex were expressed after ex-vivo cell expansion, it did not affect the potential of autologous gastric tumor cell recognition. The CD94/NKG2A complex is reported to directly inhibit NK cell cytotoxicity through recognition of HLA-E . Although both autologous gastric tumor cell lines did not express HLA-E (Table 3), we can not rule out that autologous tumor cell recognition will not be affected in HLA-E expressing tumors.
The ligands for natural cytotoxicity receptors NKp30, NKp44 and NKp46 are currently unknown. However; we postulate that at least NKp46 and NKp30 may be involved in autologous gastric tumor cell recognition since lytic activity was abrogated in the presence of blocking antibody against these receptors. Since no significant change was observed in NKG2D expression on expanded NK cells, we did not directly test the involvement of this activating receptor in autologous gastric tumor cell cytotoxicity. The fact that autologous cytotoxicity was not completely inhibited by a combination of anti-DNAM-1, NKp46, NKp44 and NKp30 may indicate that NKG2D or other unidentified receptors may also be involved. Importantly, the interaction between NK cell receptors and their ligands has recently been shown to abrogate NK cell mediated cytotoxicity of human and mouse melanoma cell lines .
Of note, both tumor cell lines also expressed high levels of Fas which is recognized to establish cell death upon interaction with its ligand, Fas-ligand . In order to test the possibility of target cell-induced killing of the expanded NK cells, all NK cells were evaluated for Fas and Fas-ligand expression before and after ex-vivo expansion. Although expanded NK cells up-regulated high levels of Fas, they did not express Fas-ligand (data not shown).
It has been suggested that in order to overcome self tolerance, multiple activating receptor-ligand interactions should be engaged . Indeed, multiple activating interactions appear to be involved in autologous cytotoxicity of tumor cells derived from patient 1 when the inhibition of cytotoxicity, in the presence of all 4 antibodies, is compared with DNAM-1 or NKp30 alone (P = 0.0356 and P = 0.0165, respectively). In contrast, no significant additional decline in autologous cytotoxicity was observed for patient 2 when cytolytic activity of all four activating receptors was compared to NKp46 alone (P = 0.7359). We postulate that these data reflect variation in expression of receptor-ligand combination in humans that are known to be operative in the control of NK cell cytotoxic activity. These variations include HLA and KIR polymorphism as well as tumor type and tumor origin (e.g. primary versus metastatic tumor cells). This is illustrated in a recent report on studies in patients with multiple myeloma  where the investigators demonstrated no specific association of autologous NK cell cytotoxicity with a single activating NK cell receptor. In fact, autologous cytotoxic effects were more likely mediated by several activating NK cell receptors which is also in agreement with a previous report  demonstrating that natural cytotoxicity of resting NK cells requires co-activation by more than one receptor.
While currently elutriated cell fraction 5 is used for monocyte and dendritic cell-based immunotherapy therapy protocols [36, 37], we demonstrated that cytolytic NK cells can be expanded from elutriated cell fractions 2, 3 and 4 regardless of the cellular content of these fractions. However, since NK cell expansion from fraction 4 failed in two out of four experiments, while expansion from PBMC and elutriated cell fractions 2 and 3 was highly successful, and considering the relative high amount of erythrocytes in fraction 2, it may be best to primarily utilize fraction 3 in NK cell expansion protocols. Of note, variability in expansion rates between donors is observed and requires further testing to determine the extent of this variation in the general population. Overall, these data provide a foundation for the large-scale generation of cytolytic NK cells from elutriated cell fractions, which could be employed alone or in combination with other cellular components such as dendritic cells for application in cellular therapy of cancer.
In summary, the large amount of cytotoxic NK cells generated by this ex-vivo expansion protocol provides the numbers of NK cells that will probably be required to be effective in the case of a large tumor burden. The ability of the expanded cells to mediate ADCC offers the possibility that their effect may be amplified if given in conjunction with a cancer cell directed mAb. An important issue to address is the ability of adoptively transferred NK cells to home and infiltrate into solid tumor tissue. Although the expanded NK cells only expressed small amounts of CD62L (data not shown), which is associated with homing into secondary tissue, we postulate that trafficking into the tumor micro-environment may be enhanced by opsonizing tumor cells with chimeric antibody. Clinical studies are needed to confirm this hypothesis, as well as to establish the therapeutic benefit of infusion of large number of ex-vivo expanded autologous NK cells.
This study is financially supported by Hasumi International Research Foundation.
CJV participated in the design of the experiments, conducted laboratory studies, prepared figures and tables and drafted the manuscript. RW established gastric tumor cell lines. SR conducted laboratory studies and assisted in the manuscript preparation. DC provided the transfected cell line and advice on NK cell expansion. KH cared for patients in the study and biopsied tissue. DLM oversaw the entirety of the project and assisted in the manuscript preparation. All authors read and approved the manuscript.


text:
class PostsController < ApplicationController
  before_action :authenticate_user!
  before_action :load_postable
  before_action :set_post, only: [:show, :edit, :update, :destroy, :like, :unlike]
  before_action :owned_post, only: [:edit, :update, :destroy]
  before_action :group_approval, only: [:create, :edit, :update, :destroy, :like, :unlike]

  def create
    @posts = Post.of_followed_users(current_user.following).order('created_at DESC').page params[:page]
    @post = @postable.posts.build(post_params)
    @post.user_id = current_user.id
    if @post.save
      flash[:success] = 'The given post has been created!'
      if @post.postable_type == 'User'
        redirect_to root_path
      else
        redirect_to @postable
      end
    else
      flash[:alert] = "The given new post couldn't be created!  Please check the form."
      redirect_to [@postable]
    end
  end

  def show
  end

  def edit
  end

  def update
    if @post.update(post_params)
      flash[:success] = 'Post updated.'
      if @post.postable_type == 'User'
        redirect_to profile_path(current_user.user_name)
      else
        redirect_to @postable
      end
    else
      flash[:alert] = 'Update failed.  Please check the form.'
      render :edit
    end
  end

  def destroy
    @post.destroy
    flash[:success] = 'The given post has been deleted.'
    redirect_to profile_path(current_user.user_name)
  end

  def like
    if @post.liked_by current_user
      if @post.user.id != current_user.id
        Notification.create(recipient: @post.user, actor: current_user, action: 'liked', notifiable: @post)
      end
      respond_to do |format|
        format.html { redirect_to :back }
        format.js
      end
    end
  end

  def unlike
    if @post.unliked_by current_user
      respond_to do |format|
        format.js
        format.html { redirect_to :back }
      end
    end
  end

  private

  def post_params
    params.require(:post).permit(:attachment, :content)
  end

  def set_post
    @post = @postable.posts.find(params[:id])
  end

  def load_postable
    @klass = [User, Group].detect { |c| params["#{c.name.underscore}_id"] }
    @postable = @klass.find(params["#{@klass.name.underscore}_id"]);
  end

  def owned_post
    if @klass.name == 'Group'
      unless current_user == @post.user || current_user.role.name == 'admin' || (current_user.group_moderator?(@post.postable.id) && !@post.user.group_moderator?(@post.postable.id))
        flash[:alert] = "That post doesn't belong to you!"
        redirect_to root_path
      end
    else
      unless current_user == @post.user || current_user.role.name == 'admin'
        flash[:alert] = "That post doesn't belong to you!"
        redirect_to root_path
      end
    end
  end

  def group_approval
    if @klass.name == 'Group'
      unless current_user.group_approved?(@postable.id)
        flash[:alert] = 'You are not eligible to post without approvals!'
        redirect_to root_path
      end
    end
  end
end

redpajama_set_name: RedPajamaGithub


text:
Posted on March 28, 2017 March 28, 2017 by Juliana Spink Mills
Cha-Cha-Cha-Changes
Oh, hey! It's another character naming post! (And here's one I made earlier… *gestures like chef on cooking show*)
This time, it's about naming difficulties I ran into while working on Heart Blade.
Changing a name after a first draft is done is always tricky. A new name can change a character in unexpected ways. But sometimes, it's unavoidable. Here are three naming hurdles I came up against while revising my novel for my publisher.
1) Sometimes the character shifts and outgrows a name. Take Alex, who first emerged from my odd little writer brain as Brother Jerome. Jerome was originally supposed to be a sort of Old Master type character. The name was perfect at the time. He's a vampire, almost 1000 years old, and he used to be a knight in the Crusades. But Jerome insisted on, well, not being Jerome. He's perpetually eighteen years old, ruggedly handsome in a shaggy blond, broad-shouldered-from-sword-work sort of way. He's covered in tattoos. And despite being an honest-to-goodness monk (though 'recently' ordained, I should add – only a couple of hundred years ago!), his penchant for wearing jeans, black tees, and an old pair of converse sneakers under his robes were a dead giveaway that I had the wrong name.
I renamed him Alexander of York and the poor guy got a whole new lease of immortal life.
2) Sometimes a character is too close to another writer's character with the same name. I had this problem with Rose, née Lila. I have big plans for Rose in book 2! She's a little edgy, and a little angry, with a lot of abandonment issues to work through. Her original name was Lila, which I loved. But then a couple of my critique partners had a Lyla in a co-authored story, and after a while their Lyla began bleeding into my Lila. They're very different characters, but there are also a few similarities, and the name just stopped working. I needed my Lila to be 100% mine. So I ditched the name. It took me forever to find a new name I liked, one that showed her as she is in Heart Blade, but could be changed slightly by Rose herself to suit who she starts to become in book 2. I won't tell you what she renames herself – you'll have to wait for Night Blade for that. But I'm happy with Rose, and I'm glad she's made the name her own.
3) Sometimes everyone just hates the name you pick! My main guy, Ash, was originally called Jimmy. It made sense to me: his full name in that first version was James Arthur Deacon III, after his father and grandfather. Jimmy matched the sweetness inside him. But although – interestingly enough – the guys who beta read the story for me were fine with the name, it got a resounding NO from all my female readers. This one took me a long while to puzzle out. I still wanted the family legacy thing to go on: Ash/Jimmy carries a pretty hefty family burden on his shoulders. So I decided to keep James Deacon and change his middle name. The men in his family would all have the same first and last names, but different middle names. The catch: it had to be a bible name. Ash's family is descended from angels and they have an important role in policing the preternatural community. I went through a gazillion naming websites before I hit on Asher, a beautiful Old Testament name that just sounded right. (Kudos to my daughter, who suggested it in the first place.) I tried it out on a few female friends and relatives and everyone agreed it was a keeper. Jimmy was out – Ash was in.
I love the three new names, and can't imagine my characters being anything else now. And the time I spent agonizing over the changes meant time spent thinking deeply about who those characters were and what really made them tick. That's the light at the end of that particular tunnel: once you find the right shiny new name, you'll feel you know your character even better than you did before.
May all your character naming problems be easy to solve! And now (because how could I not!), the gentle reminder that maybe get a second opinion if you're in doubt. Courtesy of Friends and the inimitable Phoebe Buffay.
Categoriescharacters Tagsamwriting, books, character names, characters, editing, inspiration, novel, revision, writing, YA
2 Replies to "Cha-Cha-Cha-Changes"
Em says:
I much prefer Jimmy to Ash (I am biased though – go read Otherworld…) I'm 25% through now, really enjoying it. Alex is always the best name for handsome males, I find.
I'd forgotten he was a Jim! I remembered him as James… Otherworld is great! I'm glad you're enjoying Heart Blade. 🙂
Previous PostPrevious Read For Pixels 2017 – International Women's Day Edition
Next PostNext Heart Blade Sale Week


This is a standard curriculum used in many school districts. D'Nealian can be challenging for children with handwriting difficulties for a couple reasons. The children are taught to write letters with a slant. In addition, "monkey tails" added to the manuscript or printed letter. This ending stroke is used later as the connector in cursive writing. Common modifications for children struggling with this curriculum are to omit the slant and leave off the "monkey tails" on manuscript letters.
"The First Strokes Multi-Sensory Handwriting Program was designed by an occupational therapist to teach print handwriting on a multi-sensory basis. Inherent in the program, is large gross-motor (kinesthetic) learning of the lower and upper case, print letters." Billy the Seal is also used to engage children in their learning.
A developmental program that relies heavily on multi-sensory teaching methods which work well for typically developing children, along with children with special needs. Brief but effective practice sessions are used. Read reviews.


Working again in the Philadelphia Airport, Russ Kelly Inc. had to face deal with some difficult logistics as they were working overtop of the US Airways main baggage claim operations. The storefront had to be built with precision so each corian panel would fit. This was important due to the fact that the lighting diffusers and ductwork needed to fit into the ceiling in a very small space.
High end mill work was done to complete a clean and elegant store at the Philadelphia Airport. This included a pure white corian storefront, white lacquer finished cabinets with LED lighting, and an ornate hanging chandelier in the center of the store.


text:
Jony and Marc's
(RED) Auction
An Exact Replica of Jimmy's Lambretta from the film "Quadrophenia"
30,000 — 50,000
LOT SOLD.	209,000 USD
Jony and Marc's<br>(RED) Auction
23 November 2013 | 7:00 PM EST
Chairman, Sotheby's Europe
oliver.barker@sothebys.com
Jodi Pollack
Co-Worldwide Head of
20th Century Design
jodi.pollack@sothebys.com
Elizabeth Vasold
Valuations & Estate Management, New York
elizabeth.vasold@sothebys.com
Signed by Pete Townsend and Roger Daltrey for the (RED) Auction 2013.
Metal, rubber, leather, aluminium and mirror
55 x 75 x 32 1/2 inches (140 x 190 x 83 cm)
Produced by David John Wyburn, United Kingdom
Catalogue Note
Re-sprayed in traditional Lambretta paint celly and additional paint as per the "Quadrophenia" film spec. The scooter comes fitted with rare original parts including: 1978 Southend Shield Badge, 1970's Lambretta Wassel Badge As Film Not A Repro, Original Font 1970's Plates (taken from the film original), Stadium Mirrors as per film, Original 1970s Pirelli Motor Scooter Tyre (not for road us), Michellin Acs 70s Tyre (not for road use), Rare Original Vigan Red Gem Fork Covers as per film, Original Spec Screen with all film stickers reproduced to the original spec including actor's protection strip on the top of the screen, Original Ken Cobbin Air Canister as per film, Horns and Nos Seat (Gaman) as per film, Outrigger Exhaust as per film. Jimmy's iconic Lambretta in "Quadrophenia" was a rare late 1967 LI Model. The replica scooter in the auction is an F Reg 1967.
David John Wyburn, a builder from Southwest England, has been hailed for his remarkable scooter replicas, earning him the honorific "The Oracle of Quadrophenia." He considers this Lambretta among his best work, and Jony and Marc are plainly smitten with this creation, which is both a celebration of pop culture and a powerful reminder of "Quadrophenia," a film that played a formative role as they were growing up and experimenting with design. More than just a means of transportation, the scooter represents reaction and rebellion, fundamental motivations for design. Like the Harley Davidson "Captain America" chopper from "Easy Rider," this scooter is a model of customisation, an important form of popular design and one with tremendous resonance for Jony and Marc: both of them spent much of their childhood tinkering away in the garage.
Estimate 30,000 — 50,000
Lambretta presents as a very fresh creation which has not been used. It is finished in red, white, and blue and bears the signatures of Pete Townsend and Roger Daltrey. The seat cushion and seat back are upholstered with black vinyl, and there is a contoured black rubber floor mat with red highlights. The signatures are not damaged or rubbed, and the whole of the exterior is fresh and unworn, including the red faux leopard-skin spare tire cover, chrome adornments, and raccoon tail on the rear mast. The engine appears to be complete and functional with no sign of damage or major wear from use, and the tires appear to be unworn as well. This piece was obviously purpose-built for display only, and it has been treated accordingly. Overall very good condition. This work was selected by Jony Ive and Marc Newson for the (RED) 2013 auction. This scooter includes a number of original vintage elements that show gentle age and use. Please see the catalogue for the full list of original elements. The windshield with a few shallow surface scratch. The seat with some light scratches and impressions from use. With minor wear to the tires. This work is being sold with a certificate from the (RED) 2013 auction.
In response to your inquiry, we are pleased to provide you with a general report of the condition of the property described above. Since we are not professional conservators or restorers, we urge you to consult with a restorer or conservator of your choice who will be better able to provide a detailed, professional report. Prospective buyers should inspect each lot to satisfy themselves as to condition and must understand that any statement made by Sotheby's is merely a subjective qualified opinion.
NOTWITHSTANDING THIS REPORT OR ANY DISCUSSIONS CONCERNING CONDITION OF A LOT, ALL LOTS ARE OFFERED AND SOLD "AS IS" IN ACCORDANCE WITH THE CONDITIONS OF SALE PRINTED IN THE CATALOGUE.


Next up is Exit Festival in Novi Sad, Serbia and Montenegro.
BBC Radio1 is going to broadcast from there. On that site it says that there will be an interview with Underworld and the concert. Lets hope there's more then only a few highlights of it broadcasted.


text:
Do Britain's super-wealthy non-doms bring in huge tax revenues?
September 9, 2014 by Nick Shaxson Leave a Comment
Britain's antiquated "non-domicile" rule is a relic from an age of empire. It involves defining a person's tax residence according to, in essence, their tribal affiliation. It's not the same as nationality or tax residence. As the UK government defines it:
"broadly speaking you have your domicile in the country that is your 'real' or permanent home which, if you have left, you intend to return to."
So people with some deep-rooted allegiance elsewhere (or so they might claim) can live in the UK and enjoy all its public and private services and amenities, and yet be treated as if they are not tax resident in the UK like everyone else who enjoys and pays for those same amenities. If you are a 'non-dom' then (to over-simplify) you are taxed only on the income earned domestically, and not on your income earned overseas. Of course, if you're a wealthy person too, the trick then is to shift all your income overseas so you escape tax altogether.
One set of rules for the wealthy, another set for the rest of the UK's citizens – and it also means that a lot of people who would live in other (often poor) countries and pay taxes there, live in London and effectively pay tax nowhere. All this is profoundly damaging, as we've remarked before.
"Ah, but!" cry the defenders of non-dommery. "Just look at all the tax revenues that the non-dommers pay in the UK! Don't kill the golden goose!."
And it's true – in so far as it goes. Private Eye reports:
"Latest figures, for 2011/12, show that non-doms paid £6.3 billion income tax, which sounds a fair bit and is used by tax advisers to support the "golden goose" argument: that if the non-doms were fully taxed they would stop paying this useful amount."
But here's the catch. There are non-doms and there are super-wealthy non-doms. Private Eye continues:
"What they don't say is that non-doms also paid £2.1bn in national insurance, proving that the money comes not from the entrepreneurs and ultra-wealthy who dominate the top of the rich lists, but mainly from those earning salaries from UK jobs, largely bankers but not the footloose ultra-wealthy – who, while they buy up the British property market, evidently pay little income tax."
Martin Wolf, the Financial Times' chief economics correspondent, pursued the non-dom logic, reminding us that the race to the bottom in getting poorer people to subsidise the super-rich does not stop when their tax reaches zero: the logic of the race turns it negative.
"Non-doms, we are told, make a gigantic contribution to the economy. If they are taxed too heavily, they will depart and the economy will suffer. Again, why not pursue this argument a little further? Should the UK not subsidise the inflow of human capital, just as many countries subsidise inflow of foreign direct investment? What about a negative tax (a subsidy) on all UK income earned by non-doms above, say, £100,000 a year?
Yet this, too, ought to be extended to highly paid citizens who, presumably, also provide big benefits to the economy. Why should the country wish to subsidise people to employ foreigners instead of citizens. So why not give everybody who earns about £100,000 a year a negative tax rate or at least a nice juicy lump sum?"
This reveals the nonsense at the heart of the UK vision of 'competitiveness', where the poor subsidise the rich in order to win some sort of 'global race.'
Also note that those salaried people who are paying the lion's share of the non-doms' six billion pounds' worth of taxes, as we well know, don't up sticks when asked to pay tax (see here, here or here – just for instance; with evidence that the top destination for people who flee the UK is . . . high-tax France.)
And let's be honest, the lion's share of those super-wealthy non-doms are predominantly in London, and for mostly non-tax reasons: that it's a global city with fabulous schools and amenities, where they can do their deals and speak English, and so on. Tax is a factor, but at the margin.
So what this non-dom tax system is doing is reaping dubious peanuts in terms of the relatively small numbers of houses they buy and other bits and pieces – which they may well own even if they weren't resident in Britain – but at an enormous tax cost, not to mention the costs to democracy and the rule of law.
And that's what being 'competitive' has come to mean in modern Britain.
The Spider's Web: Britain's Second Empire – released for home viewing
Quote of the day – London and money laundering
Stiglitz to tax haven UK: you are aiding and abetting theft
nine − = 5


The act of suicide is a powerful way of sending a message. Unfortunately for the victim, it's the final message. And, even more unfortunate for the survivors, it's a message filled with ambiguity that will be interpreted and re-interpreted perhaps for an entire lifetime. That burden on those left behind has led some to theorize that suicide is actually a form of homicide. It can wipe out an entire family. The legacy of multiple passive suicides did mine.
Therefore, there are probably too many at Kilpatrick Stockton preoccupied with the Mark Levy suicide this morning. Levy shot himself in the head at the office, not at home, not in a hotel, not in the woods. To me that's a type of performance art. It's meant to communicate a message.
What message could Levy have been sending? Could that message have been sent in a way that hurt the survivors less?
The use of contingency agreements between government entities and private counsel has always been controversial.
In the 1980s, the matter came before the California Supreme Court in a public nuisance case. The CA SC ruled against contingency. Then the issue emerged again in the Santa Clara lead paint public nuisance lawsuit. The defendants pushed against the use of contingency. The lower court agreed. The appeals court did not, okaying it as long as the government entity maintains full control. Here is that appeals court opinion - pay close attention to the concurring one Download H031540. No way can that control being maintained, decried the defendants. They requested and got a review by the CA SC.
Since then, the contingency debate has turned red hot. In Pennsylvania there's a motion for that state's SC to review the alleged link between awarding contingency contracts, without competitive bidding, to major campaign contributors. In Florida is a bill to make transparent every aspect of that process in the office of the attorney general. Here is the opinion of Lisa Rickard, president of the Institute for Legal Reform, on that Sunshine legislation. As may of you know, the Rhode Island SC okayed the use of contingency Download Statev.LeadIndustriesAssoc.,Inc.
That's where things stand now. Given the influence of the CA SC rulings, the decision could impact everything from the power of the state attorneys general to the number of class-action suits filed. Amicus briefs have been submitted to the CA SC. One has been filed by the National Paint & Coatings Association. As you might recall NPCA had submitted a seminal amicus brief to the RI SC in the lead paint public nuisance litigation review Download RIamicusbrief. As I saw it, the NPCA brief provided the RI High Court with a narrow passageway through the nine years of legal proceedings in the litigation. The Court overturned the verdict against the defendants.
The author of both the CA SC amicus brief and that RI one is Thomas J. Graves, Vice President and General Counsel at NPCA. Both briefs were prepared by appellate attorney Eric Lasker of Spriggs & Hollingsworth, Wahington DC.
In this amicus brief filed by NPCA, Graves asks the CA SC to reverse the appellate decision which would permit state and local entities to enter into contingency fee agreements with private attorneys. Here is a copy of the NAPA brief Download AmicusBrief.
In this particular situation, Graves goes on to point out, the agreements involve pursuing public nuisance suits based on the premise that the government lawyers can remain neutral and supervise these "subordinate" private attorneys. That would, it's contended by contingency supporters, prevent any pecuniary interests tainting the impartial pursuit of justice.
NPCA, notes Graves, concurs with the defendants that the CA SC case precedent deems these types of arrangements as anathema to ensuring the government lawyer's neutrality, which is essential to the system of justice and the confidence of society in that system.
By allowing private attorneys, continues the brief, to take this case on behalf of municipalities against old lead paint and pigment manufacturers, NPCA emphasizes, the Court of Appeal failed to recognize the distorting impacts of contingency fee arrangements. Those include not only the effect on the decision-making of the private attorneys retained but also on the decision-making of the government attorneys who retained them. At stake is the proper balancing of government authority.
Perhaps Graves's strongest point is that in practical reality, the government has an artificial incentive to pursue monetary damages award (in the billions) rather than to seek purely equitable or injunctive relief (or electing to suspend the litigation in favor of other government public interest action.) The other options may not be all that attractive because they don't provide the financial payout the government now needs to retain its outside legal team.
Thus, summarizes Graves, the enticement of a "no-cost" arrangement actually comes at the price of tipping the scales in the balancing of interests in the abatement of public nuisances of the sort the CA SC has already pronounced "cannot be tolerated."
At the National Paint and Coatings Association, Thomas J. Graves, General Counsel, can be reached at TGraves@paint.org, 202-462-8743.
Letting the sun shine in isn't always easy, particularly politically. Here, Lisa Rickard, president of the Institute for Legal Reform, deconstructs the benefits which would come from the Florida bill bringing transparency to the state attorney general's office as well as the obstacles.
The gains would include preventing those pay-to-play scandals as well as needless lawsuits. The obstacles, you got it, are being put in place by those who are gaining currently from those deals with government.
A hopeful sign, points out Rickard, Florida's own Attorney General "McCollum has championed this legislation that would require him and future Florida AGs to do the public's business, believe it or not, in public."
Twice "Without a Trace" had episodes exploring the too-high cost we human beings allow ourselves to pay for our careers. Because of that tragic flaw, Agent Jack Malone was put in mandatory therapy. This week a wounded journalist tackles with the same demon.
At Kilpatrick Stockton a lawyer might have paid the ultimate price. Today, Marisa McQuilken reports in LEGAL TIMES that there was a death of an attorney at law firm Kilpatrick Stockton. [Yesterday, the firm laid off 24 associates and counsel.] There is no comment from the firm but McQuilken tells us that Kilpatrick Stockton "posted an internal e-mail sent to firm employees Thursday morning, stating that the Metropolitan Police Department was responding to 'an unconscious male with a gunshot wound to the head on the 11th floor.'"
Unemployed attorneys - there's hope and help. Free one-hour presentation/career transition coaching by email [Mgenova981@aol.com] or phone [203-468-8579]. You have nothing to lose by contacting Jane Genova. Here's my own story of a career which had cost me way too much, including my talent Download Geezerguts.
"Resilience" by Elizabeth Edwards - Will this bring back career of John Edwards?
Part of her new book "Resilience" is about forgiveness, says Elizabeth Edwards. Well, good for her. We who have done our time in anything from 12-step programs to Buddhist monasteries know the power of letting go of resentments. It the process works, we're free of all that.
But, my experience with Elizabeth leads me to see this book as her attempt to catapult a comeback for her husband John and the father of her children. During the husband's presidential campaign I spoke with Elizabeth about items I had posted on my communications blog. She was no ordinary political wife. Cool as a cucumber but warm, highly intelligent but operating on a platform of common sense, Elizabeth is the most persuasive influential I have ever encountered.
It's in her family's self-interest that John restores his good name and puts his talents to good use in public service. This book could jump-start that. And why not let the miscreant have a second shot? After Eliot Spitzer is on his redemption stump, Henry Blodget is doing just fine again, and Larry Summers is back in the big time.
Was the supposed non-announcement that Rhode Island former U.S. Senator Senator Lincoln Chafee was running for governor meant as a trial balloon? If so, it crashed to earth with a thud. And his opponent or opponents can make plenty of "Saturday Night Live" type humor of this. I would.
Today, in THE PROVIDENCE JOURNAL, Amanda Milkovits and Katherine Gregg report that Chafee has said "that his interview on MSNBC's Rachel Maddow Show Tuesday night wasn't intended to be an 'official announcement' as a candidate." He added that he "was bolstered by the 'encouragement' he's gotten during his exploratory campaign."
This guy has been in politics. And in the tough political state of RI. His father had been a senator and a RI governor. He knows the game. He knows how to word statements so that they don't mislead. So, what's the story, former U.S. Senator Chafee?
The amazing thing is that employees - full-time, part-time, just-in-time - would have any personal information or item in the digital system of the organization which signs their check. More than two decades ago we were wise to the folly of that when, at Kraft in Rye Brook, New York, we went through the network and find a higher-up's resume, along with a cover letter. When I consult onsite, you bet I bring my own digital equipment for my communications with other accounts and friends.
So, why is there an article today by Joseph De Avila in THE WALL STREET JOURNAL about the terminated being stunned that their "digital belongings are company property?" As De Avil reports, "Employees who spend the majority of their time at the office often treat the company PC as their personal gadget, filling it with music, photos, personal contacts."
Given that we live in a litigious era, this mixing of company with self is especially dumb. Anything with an electronic trail can be used against an employee, including the allegation of excessive amount of time spent on personal communications.
The imperative to separate work for an organization from all-else might extend to other aspects of communications such as even elevator talk. That's exactly why social or emotional intelligence, particularly about being smoothly guarded, has become such a survival skill. Here's one e-book on that Download CUsersjasneDocumentsjg and here is another Download SavingSoulsJaneGenova.
Politics could take a major setback in how decisions, such as contingency agreements, are made in the office of the attorney general. Often, as we experienced in Rhode Island lead paint public nuisance litigation, contingency contracts are awarded by the AG without competitive bidding or much if any transparency. Florida could lead the way on this movement to opening up the doors to the powerful function of the state AG.
In FL, a bill is waiting the approval of House Speaker Larry Creful. It would require, reports John O'Brien in LEGAL NEWSLINE, "more transparency in the Florida Attorney General's office ... Among the regulations set forth are a cap on attorney's fees in contingency fee cases, the requirement of time sheets to determine how much work the outside counsel performed, the availability in the public of any contingency fee contract and a document signed by the attorney general every time outside counsel is hired stating its help is essential to winning the case."
The original bill made it through the House but was amended in the Senate and then passed.
Journalist Gregg Braden says that cosmic shift scheduled for 2012 will happen because of "a reversal of the Earth's magnetic poles." For BigLaw, it is appearing that what's being referred to as "2012" is already under way. No, in the legal business, this is no ordinary downturn. This is a systemic and irreversible upheaval.
Take breaking news about Cravath, Swaine & Moore. As Susan Beck reports in THE AMERICAN LAWYER, this once pillar of the White-Shoe Legal Establishment experienced a 13% drop in revenue and a 24% decline in PPP. Its cash cow - that deep-pocketed Wall Street - is no more, at least not for a long time. Beck observes that "the future" of Cravath" and about 14 other elite firms "is as uncertain as the Wall Street world they draw their strength from."
The layoffs go on and on. The latest are the 10 more associates cut at Wildman, Harrold, Allen & Dixon. Nixon Peabody reduced first-year salaries to $145,000 from $160,000. Meanwhile associates at Nixon Peabody are bumping into Pay-for-Performance slashes of up to 20 percent in salary.
What's going down, pun intended, in BigLaw is analogous to what occurred in the early 1980s in Corporate America. That whopper of a recession, along with the emergence of a global economy driven by technology, triggered a model restructuring which is still a work-in-progress. The term "redundancy" was shorthand for radically eliminating full-time high paid employees who had benefits.
A pioneer in this former GE Chief Executive Executive Jack Welch was nicknamed Neutron Jack. After he visited a GE facility, the buildings were left but the people were disappeared. The new workforce became just-in-time, were paid standardized contract wages, and had no benefits. Management visionary Charles Handy described this emerging model in his book "Age of Unreason." Incidentally, those good corporate jobs lost today are never coming back. That work has been re-distributed among just-in-time workers like myself.
The current economic crisis is plenty of reason as well as an excuse for law firms to make urgency reductions-in-staff. But you can bet they're calling in the consultants on how to restructure, particularly since, as Beck notes, even the deepest pockets are sending letters demanding, not requesting, discounts in their bills. Often the General Counsel has to get approval for expenditures from Central Purchasing. The spending leash is a lot shorter.
In addition, we all know about technology. Cloud computing, for instance, makes much of law more portable. That makes it more possible and lower cost for lawyers to start their own boutiques. And those boutiques, like The Rimon Group, can be virtual. Virtual means billing at significantly less than what BigLaw has to charge because of overhead.
Then, of course, there are all those model deletions and additions involving outsourcing, automation, and doing more assignments in-house.
Prediction: Law will be not for those who want to become wealthy but, like writing, for those who love the craft. Some anticipate starting salaries in larger law firms at about $75,000.
Already 20,000 attorneys are stranded in unemployment without a port to land in. There will probably be more, predicts the Am Law Report. BigLaw overhired without comparable growth in revenues or PPP. Now, with demand down and no uptick in the near future, those who lose jobs will have to think how to leverage their knowledge, experience, skills outside law-firm life.
Thus we are seeing accidental entrepreneurs among former risk-averse lawyers. No surprise, with student loans to still pay off, they are focused on enterprises with low or no startup costs. That brings them to the web. One caution there. We digitals are still struggling with how to monetize our ventures in cyberspace. And one thing has recently become clear. That stream of revenue ain't going to come through selling advertising.
First of all advertising is dying in a era when the tools exist to promote ourselves, our brandnames, our services and products without the advertising middleman. Secondly, advertising never had much credibility. Now with the supposed authenticity of the web, it has even less. Third, it's expensive. Few have the money to risk on taking out an ad.
In the May 11, 2009 edition of FORBES, Rebecca Buckman reports on how the smart money, including that being invested by venture capital firms, is shunning anything that runs on ads. She quotes Robert Ackerman, who is with Allegis Capital, as observing, "Nobody wants to hear about ad-supported models."
This migration away from advertising isn't just playing out in new economy businesses. Old economy empires are collapsing because they were built on ads. Think Conde Nast. PORTFOLIO just folded. And its other titles aren't doing too hot either. As THE NEW YORK POST notes, ad pages are down 31% at iconic VOGUE and 49% at ARCHITECTURAL DIGEST.
So, how do we monetize our web enterprise?
An excellent model is Mediabistro.com, which was sold by the founder for multi-millions. It provides special events, training seminars, access to affordable health and dental benefits for members, classified, and a sense of community.
Another viable model is using the site as a primary new-business development tool. That's how I monetize - indirectly - this and my two other - here and here - blogs.
A third model is the site as a platform for the social marketing of products and services. It's an easier sell because that's how social media work.
Forget ads? The display kind, probably. But classified still can pull in revenue.
A useful read might be the 2008 guide "The Truth about Profiting from Social Networking," by Patrice-Anne Rutledge.